WO2015148648A1 - Formulation d'adjuvant non toxique comprenant une composition de liposomes contenant un lipide a monophosphorylé (mpla) et une saponine - Google Patents

Formulation d'adjuvant non toxique comprenant une composition de liposomes contenant un lipide a monophosphorylé (mpla) et une saponine Download PDF

Info

Publication number
WO2015148648A1
WO2015148648A1 PCT/US2015/022461 US2015022461W WO2015148648A1 WO 2015148648 A1 WO2015148648 A1 WO 2015148648A1 US 2015022461 W US2015022461 W US 2015022461W WO 2015148648 A1 WO2015148648 A1 WO 2015148648A1
Authority
WO
WIPO (PCT)
Prior art keywords
less
mol
mpla
adjuvant formulation
saponin
Prior art date
Application number
PCT/US2015/022461
Other languages
English (en)
Inventor
Carl R. Alving
Zoltan Beck
Original Assignee
The Government Of The United States Of America As Represented By The Secretary Of The Army
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The Government Of The United States Of America As Represented By The Secretary Of The Army filed Critical The Government Of The United States Of America As Represented By The Secretary Of The Army
Priority to EP15716264.5A priority Critical patent/EP3122380A1/fr
Priority to JP2017502920A priority patent/JP6608422B2/ja
Priority to AU2015236106A priority patent/AU2015236106A1/en
Priority to RU2016141622A priority patent/RU2016141622A/ru
Priority to CA2943190A priority patent/CA2943190A1/fr
Priority to MX2016012168A priority patent/MX2016012168A/es
Priority to KR1020167026025A priority patent/KR102242875B1/ko
Priority to SG11201607396WA priority patent/SG11201607396WA/en
Priority to US15/127,081 priority patent/US10434167B2/en
Priority to CN201580026334.9A priority patent/CN107124869B/zh
Publication of WO2015148648A1 publication Critical patent/WO2015148648A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A61K39/21—Retroviridae, e.g. equine infectious anemia virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55555—Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55572—Lipopolysaccharides; Lipid A; Monophosphoryl lipid A
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55577—Saponins; Quil A; QS21; ISCOMS
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16171—Demonstrated in vivo effect

Definitions

  • compositions comprising a monophosphoryl lipid A (MPLA)-containing liposome and at least one saponin, e.g., QS-21.
  • the liposome comprises i) a lipid bilayer comprising phospholipids in which the hydrocarbon chains have a melting temperature in water of > 23 °C and ii) cholesterol at a mole percent concentration of greater than about 50% (mol/mol).
  • LPS Lipopolysaccharides
  • lipid A which is the innermost region of LPS that serves as the hydrophobic anchor and comprises phosphorylated glucosamine disaccharide units carrying long chain fatty acids.
  • LPS lethal toxicity, pyrogenicity, and adjuvanticity
  • lipid A moiety of lipid A
  • Both LPS and lipid A have long been known for their strong adjuvant effects, but the high toxicity of these molecules has limited their use in vaccine formulations.
  • Significant scientific effort has occurred towards reducing the toxicity of LPS or lipid A while maintaining their adjuvanticity.
  • Quillaja saponins are a mixture of amphipathic triterpene glycosides extracted from the bark of the tree Quillaja saponaria. Crude saponins have been employed as veterinary adjuvants.
  • Quil-A is a partially purified aqueous extract of the Quillaja saponin material
  • QS-21 is an HPLC purified fraction of Quil A. Methods of producing QS-21 (referred to as QA21 ) are disclosed in U.S. Pat. No. 5,057,540.
  • One of the major side effects of using saponins such as QS-21 is its cytotoxicity. Specifically, although the exact cytotoxic mechanism is currently unknown, saponins cause hemolysis, which is the rupturing or lysis of red blood cells.
  • an adjuvant formulation comprising a monophosphoryl lipid A (MPLA)-containing liposome (L(MPLA)) composition and a saponin, wherein the adjuvant formulation displays minimal toxicity of either lipid A or saponin.
  • MPLA monophosphoryl lipid A
  • L(MPLA) monophosphoryl lipid A
  • an adjuvant formulation comprising a
  • the liposome composition comprises i) a lipid bilayer composing phospholipids in which the hydrocarbon chains have a melting temperature in water of > 23°C and ii) cholesterol at a mole percent concentration of greater than about 50% (mol/mol).
  • the saponin may be selected from QS-7, QS- 18, QS-21 , or a mixture thereof.
  • the saponin can be QS-21 .
  • the liposome composition of the adjuvant formulation may comprise cholesterol at a mole percent concentration of about 55% to about 71% (mol/mol), or preferably about 55% (mol/mol).
  • the liposome composition of the adjuvant formulation may comprise a phosphatidylcholine (PC) selected from the group consisting of: dimyristoyl phosphatidylcholine (DMPC), dipalmitoyl phosphatidylcholine (DPPC), and distearyl phosphatidylcholine (DSPC).
  • PC phosphatidylcholine
  • DMPC dimyristoyl phosphatidylcholine
  • DPPC dipalmitoyl phosphatidylcholine
  • DSPC distearyl phosphatidylcholine
  • the liposome composition of the adjuvant formulation may comprise multi-lamellar vesicles (MLV) or small uni-lamellar vesicles (SUV), wherein small uni-lamellar vesicles are about 50 to about 100 nra in diameter, and wherein multi-lamellar vesicles are about 1 to about 4 ⁇ in diameter.
  • MLV multi-lamellar vesicles
  • SUV small uni-lamellar vesicles
  • the liposome composition of the adjuvant formulation may comprise about 5 mg or less, about 4 mg or less, about 3 mg or less, about 2 mg or less, about 1 mg or less, about 0.9 mg or less, about 0.8 mg or less, about 0.7 mg or less, about 0.6 mg or less, about 0.5 mg or less, about 0.4 mg or less, about 0.3 mg or less, about 0.2 mg or less, about 0.1 mg or less, about 0.09 mg or less, about 0.08 mg or less, about 0.07 mg or less, about 0.06 mg or less, about 0.05 mg or less, about 0.04 mg or less, about 0.03 mg or less, about 0.02 mg or less, or about 0.01 mg or less of MPLA (total weight per ml liposome suspension).
  • the liposome composition of the adjuvant formulation may have a MPLA:phospholipid mole ratio of about 1 :5.6 to about 1 :880, or about 1 :88 to about 1 :220.
  • a method of reducing toxicity of a saponin as an adjuvant or preparing an adjuvant formulation comprising adding a monophosphoryl lipid A (MPLA)-containing liposome composition to the saponin, wherein the liposome composition comprises i) a lipid bilayer comprising phospholipids in which the hydrocarbon chains have a melting temperature in water of > 23°C and ii) cholesterol at a mole percent concentration of greater than about 50% (mol/mol).
  • the saponin may be selected from the group consisting of QS-7, QS- 1 8, QS-21 , and a mixture thereof.
  • saponin can be QS-21 .
  • the liposome composition may comprise cholesterol at a mole percent concentration of about 55% to about 71 % (mol/mol), preferably, about 55% (mol/mol).
  • FIG. 1 depicts hemolytic activities of liposome compositions with various QS-21 concentrations (FIG. 1 A: 10 ⁇ QS-21 ; FIG. I B: 50 ng/ml QS-21 ) as described in Example 1 .
  • FIG. 2 depicts hemolysis of erythrocytes by QS-21 pre-incubated with liposomes as described in Example 1 .
  • QS-21 was mixed with DMPC/DMPG liposomes containing a total of 5 ⁇ of phospholipids at 22 °C.
  • the liposomes also contained 33.7, 43 , or 55 mol% Choi .
  • FIG. 4 depicts inhibition of binding of QS-21 to liposomal Choi by liposomal MPLA.
  • the experiments were conducted as described in Example 2.
  • the mean ⁇ S.D. is shown with 14 independent liposome batches of L, and with 4 independent batches of L(MPLA).
  • FIG. 5 depicts effect of phospholipid fatty acyl chain length on accessibility of liposomal Choi for binding of QS-21 .
  • the experiments were conducted as described in Example 2.
  • FIG. 5A illustrates the results of the liposomes with 33.7 mol% Choi and no MPLA.
  • FIG. 5B illustrates the results of the liposomes with 55 mol% Choi, and no MPLA.
  • FIG. 5C illustrates the results of the liposomes with 55 mol% Choi and MPLA.
  • FIG. 7 depicts suppression of LAL positivity of DMPC/DMPG/Chol/MPLA liposome by increasing liposomal Choi contents. Results of LAL binding to liposomes containing 33.7 mol% Choi are shown in FIG. 7A, while results of LAL binding to liposomes containing 55 mol% Choi are shown in FIG. 7B.
  • MPLA phospholipid ratios of 1 :88 or 1 :5.6.
  • the curves represent ALFQ in which the initial ALF comprised (from left to right): MLV with MPLA.phospholipid ratio of 1 :220; and SUV with
  • MPLA phospholipid ratios of 1 :88 and 1 :5.6, respectively.
  • FIG. 9 depicts IgG endpoint titers to gp l40.
  • Mice were immunized with ALF+CN gp l 40 (FIG. 9A) and ALFQ+CN gp l40 (FIG. 9B) containing the indicated MPLA:phospholipid molar ratios in which 20 ⁇ g of MPLA was present in each formulation.
  • Sera were from week 9 and were assayed by ELISA. Values are the mean of 6 animals/group ⁇ standard deviation.
  • F G. 10 depicts IgG subtype profiles of anti-gp l 40 antisera as described in Example 4 and Materials and Methods. Mice were immunized with ALF+gp l 40 (FIG. 10A) or ALFQ+gp l 40 (FIG. 10B). ELISA plates were coated with gpl 40. Subtype analyses were conducted and values were calculated from standard curves. Each panel represents pooled sera from one group from week 9 of immunized mice.
  • FIG. 1 1 depicts production of IFN- ⁇ (FIG. 1 1A) and IL-4 (FIG. 1 IB) by splenocytes from mice 9 weeks after primary immunization.
  • Splenocytes were plated and stimulated with gpl40 CN54 for 18 hours, and the number of cytokine producing cells were determine by ELISPOT.
  • Vertical lines represent the mean of the 6 animals/group (Na ' ive: non-immunized control mice).
  • FIG. 12 depicts neutralization of HIV by pooled antisera.
  • the vaccine formulation is indicated on the x-axis.
  • ID50 values represent 50% neutralization of pseudoviruses MW965.26PV (FIG. 12A) and GS015PV (FIG. 12B) at the indicated dilution of the serum samples from mice 9 weeks after primaiy immunization.
  • the controls were sera from non-immunized mice.
  • ID50 values ⁇ 20 were considered as background. MuLV PV was used as assay negative control (data not shown).
  • immunogen is an agent capable of inducing humoral and/or cell- mediated immune response.
  • the immunogen as described herein can be an antigen, a hapten, or an inactivated pathogen.
  • An immunogenic composition as described herein can be, for example, a vaccine formulation.
  • Liposomes refer to closed bilayer membranes containing an entrapped aqueous volume. Liposomes may also be uni-lamellar vesicles possessing a single membrane bilayer or multi-lamellar vesicles with multiple membrane bilayers, each separated from the next by an aqueous layer. The structure of the resulting membrane bilayer is such that the hydrophobic (non-polar) tails of the lipid are oriented toward the center of the bilayer while the hydrophilic (polar) heads orient towards the aqueous phase.
  • Liposomes as they are ordinarily used, consist of smectic mesophases, and can consist of either phospholipid or nonphospholipid smectic mesophases. Smectic mesophase is most accurately described by Small, HANDBOOK OF LIPID RESEARCH, Vol. 4, Plenum, NY, 1986, pp. 49-50. According to Small, "[w]hen a given molecule is heated, instead of melting directly into an isotropic liquid, it may instead pass through intermediate states called mesophases or liquid crystals, characterized by residual order in some directions but by lack of order in others ... In general, the molecules of liquid crystals are somewhat longer than they are wide and have a polar or aromatic part somewhere along the length of the molecule.
  • the molecular shape and the polar-polar, or aromatic, interaction permit the molecules to align in partially ordered arrays ... These structures characteristically occur in molecules that possess a polar group at one end.
  • Liquid crystals with long- range order in the direction of the long axis of the molecule are called smectic, layered, or lamellar liquid crystals ... In the smectic states the molecules may be in single or double layers, normal or tilted to the plane of the layer, and with frozen or melted aliphatic chains.”
  • Lipid A is a set of complex, heavily acylated and amidated diglucosamine diphosphate molecules and is the lipid moiety common to all lipopolysaccharides (LPS; also known as endotoxin) from Gram-negative bacteria.
  • LPS lipopolysaccharides
  • LPS covers virtually the entire outer surface of all Gram-negative bacteria, and lipid A anchors the LPS into the outer lipid surface of the bacterium.
  • the O-polysaccharide portion of LPS in wild-type smooth bacteria is linked to a relatively conserved core oligosaccharide that is expressed in rough mutants, and this in turn is linked to lipid A through highly conserved 2-keto-3-deoxyoctanoic acid sugars that are unique chemical structures sometimes required for bacterial viability and found only in LPS.
  • “Monophosphoryl lipid A” is a lipid A congener in which the glucosamine- 1 -phosphate group on the polar head group has been removed. Numerous congeners of MPLA also exist.
  • the "mole percent concentration of cholesterol" of a liposome composition as used herein refers to the ratio of Choktotal phospholipid (i.e., phosphatidylcholine and phosphatidylglycerol) originally used in the preparation of the liposome composition.
  • a “physiologically acceptable vehicle” as used herein refers to a vehicle that is suitable for in vivo administration (e.g., oral, transdermal or parenteral administration) or in vitro use, i.e., cell culture.
  • exemplary physiologically acceptable vehicles can be those physiologically acceptable constituents of liposomes as disclosed in U.S. Patent Nos. 4,186, 183 and 4,302,459.
  • a suitable saponin is Quil A, its derivatives thereof, or any purified component thereof (for example, QS-7, QS- 18, QS-21 , or a mixture thereof).
  • Quil A is a saponin preparation isolated from the South American tree Quillaja Saponaria Molina and was first found to have adjuvant activity.
  • Liposomes are closed bilayer membranes containing an entrapped aqueous volume. Liposomes may also be uni-lamellar vesicles possessing a single membrane bilayer or multi-lamellar vesicles with multiple membrane bilayers, each separated from the next by an aqueous layer.
  • the structure of the resulting membrane bilayer is such that the hydrophobic (non-polar) tails of the lipid are oriented toward the center of the bilayer while the hydrophilic (polar) heads orient towards the aqueous phase.
  • Suitable hydrophilic polymers for surrounding the liposomes include, without limitation, PEG, polyvinylpyrrolidone, polyvinylmethylether, polymethyloxazoline, polyethyloxazoline, polyhydroxypropyloxazoline,
  • Liposomes can be made without hydrophilic polymers. Therefore, liposome formulations may or may not contain hydrophilic polymers.
  • Liposomes may be comprised of any lipid or lipid combination known in the art.
  • the vesicle-forming lipids may be naturally-occurring or synthetic lipids, including phospholipids, such as phosphatidylcholine,
  • MPLA serves as a potent adjuvant and serves to raise the immunogenicity of the liposome and peptides, proteins, or haptens associated with the liposome.
  • a typical monophosphoryl lipid A (MPLA)- containing liposome (L(MPLA)) is the one originally referred to as Walter Reed liposomes but now known as Army Liposome Formulation (ALF), as a vaccine adjuvant.
  • MPLA monophosphoryl lipid A
  • ALF Army Liposome Formulation
  • an L(MPLA) may comprise a
  • PC phosphatidylcholine
  • DMPC dimyristoyl phosphatidylcholine
  • DPPC dipalmitoyi phosphatidylcholine
  • DSPC disteaiyl phosphatidylcholine
  • the L(MPLA) may also comprise a
  • Adjuvant formulations comprising liposome and saponin
  • the AS01 formulation is being developed as an adjuvant for a variety of vaccines. See Garcon & Mechelen, 201 1 , Expert. Rev. Vaccines, 10: 471-86.
  • the AS01 formulation as described in U.S. Published Patent Application No. 201 1/0206758, may contain cholesterol (sterol) at a mole percent concentration of 1 -50% (mol/mol), preferably 20-25% (mol/mol).
  • the adjuvant formulation comprises
  • the saponin may be selected from QS-7, QS-18, QS-21 , or a mixture thereof, or the saponin preferably may be QS-21.
  • the adjuvant formulation may contain about 1 mg or less, about 0.9 mg or less, about 0.8 mg or less, about 0.7 mg or less, about 0.6 mg or less, about 0.5 mg or less, about 0.4 mg or less, about 0.3 mg or less, about 0.2 mg or less, about 0.1 mg or less, about 0.09 mg or less, about 0.08 mg or less, about 0.07 mg or less, about 0.06 mg or less, about 0.05 mg or less, about 0.04 mg or less, about 0.03 mg or less, about 0.02 mg or less, or about 0.01 mg or less of saponin per ml liposome suspension.
  • DMEM Dulbecco's modified Eagle's medium
  • FBS fetal bovine serum
  • penicillin 100 unit/ml penicillin
  • streptomycin 100 g/ml streptomycin
  • 2 niM L-glutamine was used for culturing TZM.bl cells and for HlV- 1 neutralization assays.
  • Liposomal Choi concentration was 43 mol% in all ALF preparations lacking QS-21.
  • Liposomes were microfluidized using a Microfluidics LV 1 low volume high shear microfluidizer (ATS Scientific Inc., Burlington, Ontario, Canada) at 30,000 psi to form SUVs.
  • the diameter size distributions of liposomes were measured by Horiba LB-550 particle sizer (Horiba Scientific, New Jersey, NJ).
  • Army liposome formulations plus QS-21 (ALFQ) were made by mixing MLV or SUV liposomes in which the Choi concentration was 55 mol% with QS-21 .
  • the QS-2 1 irreversibly binds to the Choi in the liposomes under these conditions with no detectable free QS- 21 present in the buffer.
  • the total amount of QS-21 in each ALFQ formulation used for immunization was 10 ⁇ g.
  • Cholesterol content was analyzed to confirm the cholesterol, and indirectly, the phospholipid concentration of the liposome formulations by the established methods. See, e.g. , Zlatkis et al., 1953, J. Lab. Clin. Med. , 41 : 486-492. Briefly, 5 to 200 ⁇ of liposomes were added to water to a final volume of 200 ⁇ . Three ml of glacial acetic acid were added, followed by the addition of 2 ml of 0.1 % ferric chloride in glacial sulfuric acid. After mixing and then equilibrating, the samples to room temperature, absorbance was read at 560 ran. The cholesterol concentration of the liposomes was determined from a cholesterol standard curve.
  • red blood cells Hemolysis of red blood cells was used as a measure both of the relative amount of free QS-21 , and of the toxicity of QS-21 under the indicated experimental conditions.
  • Human red blood cells were purchased from the Research Blood
  • Limulus amebocyte lysate (LAL) assay was used as a probe to examine the roles of phospholipid chain length and mole fraction of Choi on the liposomal surface expression of MPLA.
  • a lysate of amebocytes from the blood of Limulus polyphemus (Atlantic horseshoe crab) containing a clotting protein is widely used as a surrogate probe for detecting the endotoxic activity of LPS or lipid A. See, e.g. , Brandenburg et al., 2009, Curr. Med. Chem., 16: 2653-2660.
  • mice Female Balb/c mice (Charles River Laboratories, Indianapolis, IN, USA) (5-6 weeks of age; 6/group) were immunized intramuscularly (IM) with 0.05 ml of the vaccines by injection in alternative rear thighs at 0, 3 and 6 weeks.
  • IM intramuscularly
  • Four mouse groups were immunized with the ALF+gp l 40 formulations listed above and three groups were immunized with the ALFQ+gp MO formulations.
  • One mouse group was not immunized, and was used as negative control for the immunological studies.
  • the animals were bled prior to the first immunization and 3 and 6 weeks after the primary immunization. At week 9, the animals were terminally bled and the spleens were collected.
  • HRP-linked sheep anti-mouse IgG (0.1 ⁇ g in 100 ⁇ blocking buffer) was added to each well and plates were incubated for 1 hour followed by washing.
  • One hundred ⁇ of ABTS 2-component substrate (Kirkegaard and Perry Laboratories, Gaithersburg, MD, USA) were added to each well, and plates were incubated for 1 hour. Color development was stopped by adding 100 ⁇ /well of 1 % SDS. The absorbance was read at 405 nm. End point titer was defined as the dilution at which the absorbance was twice background.
  • IgG subtype determination was modified from Glenn et al., 1995, Immunol. Let. 47: 73-78.
  • a standard curve was made by using unconjugated goat anti-mouse IgG (Fab).
  • Fab unconjugated goat anti-mouse IgG
  • One hundred ⁇ of Fab 1 ⁇ g ml in PBS was added to the wells of 96-well Immulon 2HB flat bottom plates and the plates were incubated overnight at 4°C. Plates were washed 3 times with 0.05%) Tween 20 in PBS (washing buffer), blocked with 250 ⁇ /well of 0.5%> skim milk in PBS (blocking buffer), and incubated at room temperature (RT) for 1 hour. After washing, 100 ⁇ of blocking buffer were added to each well.
  • Purified mouse IgG 1 , lgG2a, IgG2b, and IgG3 were diluted two-fold starting from 200 ng/ml of blocking buffer. One hundred ⁇ of each dilution were added to the plates. Plates were incubated at room temperature for 2 hours, and washed 3 times. One hundred ⁇ of HRP-linked goat anti-mouse IgG l , IgG2a, IgG2b, and IgG3 ( 1 : 1000 dilution in blocking buffer) were added to the corresponding wells, and plates were incubated at room temperature for 1 hour. Plates were washed and 100 ⁇ of ABTS were added. Color development was stopped with 100 ⁇ /well of 1% SDS.
  • Spleens from euthanized mice were pressed through a 100- ⁇ nylon cell strainer (Thomas Scientific, Swedesboro, NJ, USA, cat No. 4620F05) with the plunger of a syringe. Splenic cell suspensions were collected in and washed 3 times with cRPMI medium. ELISPOT was conducted as described in Rao et al., 2002, J. Virol, 76: 9176-85.
  • multiScreen 96-well microtiter plates Prior to the harvesting of spleens, multiScreen 96-well microtiter plates (EMD Millipore, Billerica, MA, USA) were pre-treated with 70% ethanol, washed 3 times with PBS and coated with 100 ⁇ /well of either 5 ⁇ g/ml of capture INF-y-specific IgG diluted in PBS or 2 igl ⁇ of capture IL-4-specific specific IgG diluted in PBS. Plates were incubated overnight at 4°C, then washed 2 times with cRPMI and blocked with 200 ⁇ /well cRMPl at RT for 2 hours. Fifty ⁇ of mouse cell suspensions (8 x 10 6 cells/ml) were plated in duplicate for each cytokine assay.
  • Neutralizing antibody responses against two tier 1 HIV- 1 Env pseudoviruses were measured by using luciferase-based virus neutralization assays with TZM.bl cells as previously described. See, e.g. , Brown et al., 2007, J. Virol , 81 : 2087-91.
  • the 50% inhibitory dilution of serum (ID50) was calculated as the serum dilution that resulted in a 50% reduction in relative luminescence units compared with the virus control wells after the subtraction of cell control relative luminescence units.
  • DMEM Dulbecco's modified Eagle's medium
  • HIV clade C, GS015 PV and MW965.26 PV (25 ⁇ ) were added to each well and the plates were incubated for 1 h at 37°C.
  • TZM.bl cells were then added ( 104 cells/well) in a 50 ⁇ volume in 1 0% DMEM containing DEAE-dextran, and neutralizing antibody titers were determined as previously described in Brown et al., 2008.
  • Murine leukemia virus (MuLV) negative controls were included in all assays.
  • the hemolytic activities of liposome compositions with QS-21 were characterized first by varying QS-21 concentration. Briefly, fifty millimolar phospholipid liposomes containing 33.7%, 43%, and 55% cholesterol were mixed and incubated with 1 0 ⁇ g/m l and 50 pg/ml QS-21 at room temperature for 20 minutes. Red blood cells were added to the mixture of liposome-QS-21 , and incubated for 30 minutes. Free QS-21 served as positive control. Cells were centrifuged and the optical density of supernatant was measured at 541 nm to reflect the levels of hemolysis. The results, as presented in FIG. 1 , suggest that the levels of hemolysis by liposome compositions with QS-21 depends upon the concentration of cholesterol in the liposomes.
  • the hemolytic activities of liposome compositions with QS-21 were further characterized by varying mol% of liposomal Choi.
  • the degree of eiythrocyte damage caused by QS-21 was found to be inversely related to the mol% of liposomal Choi that had been pre-incubated with QS-21 (FIG. 2). While free QS-21 caused maximum hemolysis at approximately 2 ⁇ g, maximum hemolysis occurred at approximately 4 ⁇ g of QS-21 when QS-21 was pre-incubated with liposomes containing either 33.7 or 43 mol% Choi (FIG. 2). Liposomes having 55 mol% Choi completely blocked the hemolytic effect of QS-21 up to 200 ⁇ g (FIG. 2). Similar results were observed for liposome compositions containing varying concentrations of saponin (FIG. 3). No hemolysis with up to 500 ⁇ g/ml saponins was detected when liposomes contained 55 mol% cholesterol.
  • LAL Limulus amebocyte lysate
  • FIG. 7A shows that the MPLA in DMPC/DMPC/Chol/MPLA, DPPC/DMPC/Chol/MPLA, or DSPC/DMPC/Chol/MPLA liposomes containing 33.7 mol % Choi was detected by the LAL assay.
  • the recognition of MPLA by LAL in liposomes containing 33.7% Choi was proportional to the length of saturated fatty acyl chains of phosphatidylcholine, with LAL binding in the order DSPC > DPPC > DMPC.
  • MLV and SUV each being a type of ALF liposomes (i.e., containing MPLA), were constructed.
  • ALF MLV was between 1 and 4 ⁇ .
  • MLV indicates that light scattering analysis detected many large particles
  • MLV also contains numerous small particles in a hyperbolic distribution as shown by electronic particle size analysis. See, e.g. , Alving et al., 1975, Biochim. Biophys. Acta., 394: 157-65.
  • FIG. 9A ALF SUV containing MPLA:phospholipid in the ratio of 1 :88 induced higher IgG anti-gpl40 binding antibody production in mice than ALF MLV having the same MPLA:phospholipid ratio in a statistically significant fashion.
  • ALF SUV with an MPLA:phospholipid ratio of 1 :5.6 induced the highest titer of binding antibodies.
  • FIG. 9B shows that all ALFQ-adjuvanted formulations induced significantly higher binding antibodies to gpl40 than any of the ALF MLV- adjuvanted formulations. The total gpl40-specific IgG binding antibody levels in all ALFQ-adjuvanted groups appeared independent of the size of the liposomes or the relative phospholipid concentration.
  • IgG subtype determination was conducted to further characterize the antibody production. All mouse groups immunized with ALF + gpl40 revealed moderate or high titers of IgG 1 binding antibodies to gpHO (FIG. 10A). ALF MLV induced dominantly IgGl subclass, while ALF SUV induced a balance between IgGl and IgG2a production and also induced IgG2b production. Increasing the expression
  • FIG. 10A As shown in FIG. 10B, in the ALFQ groups the serum concentration of IgGl was constantly high; and all subtypes were induced, but these were independent of the MPLAiphospholipid ratios of ALF SUV or ALF MLV initially utilized for creation of ALFQ.
  • ELISPOT analyses were performed on freshly isolated splenic lymphocytes from mice immunized with adjuvanted gpl40. As shown in FIG. 1 1, all of the ALF-adjuvanted formulations induced proliferation of I F-y-producing splenic lymphocytes that was higher than observed with the na ' fve control. ALFQ formulations induced significantly higher numbers of INF-y-positive cells than ALF (FIG. 1 1A). All of the immunized mice exhibited high IL-4 production that was significantly higher (p ⁇ 0.0001) than naive mice (FIG. 1 IB). Significant differences of IL-4 producing cell numbers were not observed among immunized groups.
  • the ALFQ liposomes caused a distinctive induction of INF- ⁇ secretion that was independent of the MPLA:phospholipid ratio.
  • the ALFQ liposomes might be a potentially useful adjuvant for induction of Thl -type immunity.
  • ALF having MPLA:phospholipid ratio of 1 :5.6 induced the highest neutralization titer, while ALFQ with 1 :88 and 1 :220 MPLA:phospholipid ratios also induced neutralizing antibodies, but with somewhat lower titers.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Virology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dispersion Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Communicable Diseases (AREA)
  • Hematology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Medicinal Preparation (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

La présente invention concerne une formulation d'adjuvant comprenant une composition de liposomes contenant un lipide A monophosphorylé (MPLA) et une saponine (par exemple, la QS-21), la composition de liposomes comprenant i) une bicouche lipidique comprenant des phospholipides dans laquelle les chaînes hydrocarbonées possèdent une température de fusion dans l'eau ≥ 23 °C et ii) du cholestérol à une concentration en pour cent en moles supérieure à environ 50 % (mole/mole), de préférence d'environ 55 % à environ 71 % (mole/mole), ou plus préférablement d'environ 55 % (mole/mole). La formulation d'adjuvant présente une toxicité minimale soit de la saponine soit du lipide A.
PCT/US2015/022461 2014-03-25 2015-03-25 Formulation d'adjuvant non toxique comprenant une composition de liposomes contenant un lipide a monophosphorylé (mpla) et une saponine WO2015148648A1 (fr)

Priority Applications (10)

Application Number Priority Date Filing Date Title
EP15716264.5A EP3122380A1 (fr) 2014-03-25 2015-03-25 Formulation d'adjuvant non toxique comprenant une composition de liposomes contenant un lipide a monophosphorylé (mpla) et une saponine
JP2017502920A JP6608422B2 (ja) 2014-03-25 2015-03-25 モノホスホリルリピッドａ（ｍｐｌａ）含有リポソーム組成物およびサポニンを含む非毒性アジュバント製剤
AU2015236106A AU2015236106A1 (en) 2014-03-25 2015-03-25 Non-toxic adjuvant formulation comprising a monophosphoryl lipid a (MPLA)-containing liposome composition and a saponin
RU2016141622A RU2016141622A (ru) 2014-03-25 2015-03-25 Нетоксичный адъювантный состав, содержащий композицию содержащей монофосфорил-липид а (mpla) липосомы и сапонин
CA2943190A CA2943190A1 (fr) 2014-03-25 2015-03-25 Formulation d'adjuvant non toxique comprenant une composition de liposomes contenant un lipide a monophosphoryle (mpla) et une saponine
MX2016012168A MX2016012168A (es) 2014-03-25 2015-03-25 Formulacion adyuvante no toxica que comprende una composicion de liposoma que contiene monofosforil-lipido a (mpla) y una saponina.
KR1020167026025A KR102242875B1 (ko) 2014-03-25 2015-03-25 모노포스포릴 지질 a (mpla)-함유 리포솜 조성물 및 사포닌을 포함하는 무독성 아주반트 제제
SG11201607396WA SG11201607396WA (en) 2014-03-25 2015-03-25 Non-toxic adjuvant formulation comprising a monophosphoryl lipid a (mpla)-containing liposome composition and a saponin
US15/127,081 US10434167B2 (en) 2014-03-25 2015-03-25 Non-toxic adjuvant formulation comprising a monophosphoryl lipid A (MPLA)-containing liposome composition and a saponin
CN201580026334.9A CN107124869B (zh) 2014-03-25 2015-03-25 包含含有单磷酰脂质a（mpla）的脂质体组合物和皂苷的无毒佐剂制剂

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201461970118P 2014-03-25 2014-03-25
US61/970,118 2014-03-25

Publications (1)

Publication Number Publication Date
WO2015148648A1 true WO2015148648A1 (fr) 2015-10-01

Family

ID=52829375

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2015/022461 WO2015148648A1 (fr) 2014-03-25 2015-03-25 Formulation d'adjuvant non toxique comprenant une composition de liposomes contenant un lipide a monophosphorylé (mpla) et une saponine

Country Status (11)

Country Link
US (1) US10434167B2 (fr)
EP (1) EP3122380A1 (fr)
JP (1) JP6608422B2 (fr)
KR (1) KR102242875B1 (fr)
CN (1) CN107124869B (fr)
AU (1) AU2015236106A1 (fr)
CA (1) CA2943190A1 (fr)
MX (1) MX2016012168A (fr)
RU (1) RU2016141622A (fr)
SG (2) SG11201607396WA (fr)
WO (1) WO2015148648A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017153993A1 (fr) * 2016-03-09 2017-09-14 Technion Research And Development Foundation Ltd. Formulations liposomales et leurs procédés d'utilisation en agriculture
WO2019051149A1 (fr) * 2017-09-08 2019-03-14 Infectious Disease Research Institute Formulations liposomales comprenant de la saponine et procédés d'utilisation
EP3615005A4 (fr) * 2017-04-28 2021-03-10 The Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc. Compositions et méthodes d'administration de vaccins
WO2023175454A1 (fr) 2022-03-14 2023-09-21 Pfizer Inc. Procédés de production d'adjuvant

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP6661454B2 (ja) * 2016-04-21 2020-03-11 株式会社東海理化電機製作所 表示装置、表示板の製造方法
CN107669692B (zh) * 2017-09-07 2020-09-29 中国人民解放军第二军医大学 Mpla在制备电离辐射致肠道损伤防治药物中的应用
WO2020243166A1 (fr) * 2019-05-28 2020-12-03 The Regents Of The University Of California Adjuvants de polarisation th1 pour améliorer l'immunogénicité des antigènes du vih
CN110613844B (zh) * 2019-10-23 2024-02-27 中国医学科学院生物医学工程研究所 一种迷你联合佐剂纳米颗粒及其制备方法和应用
CA3161668A1 (fr) 2019-12-20 2021-06-24 Nammi Therapeutics, Inc. Compositions liposomales formulees et/ou co-formulees contenant des promedicaments agonistes de recepteurs de type toll (« tlr ») utiles dans le traitement du cancer et methodes a ssociees
WO2022003443A1 (fr) * 2020-06-30 2022-01-06 아이진 주식회사 Composition d'inhibition de l'hémolyse induite par la saponine, contenant un liposome cationique
GB2600468A (en) 2020-10-30 2022-05-04 Excivion Ltd Adjuvant composition
CN112546233A (zh) * 2020-12-25 2021-03-26 常州大学 一种负载水不溶性免疫调节分子的透明质酸复合材料及其制备方法
CN114366810A (zh) * 2022-02-15 2022-04-19 中国人民解放军陆军军医大学 一种可高效诱导体液和细胞免疫的天然微生物和植物来源复合双佐剂及其制备方法和应用
WO2024069420A2 (fr) 2022-09-29 2024-04-04 Pfizer Inc. Compositions immunogènes comprenant un trimère de protéine f rsv
CN117771361A (zh) * 2024-02-27 2024-03-29 天津中逸安健生物科技有限公司 一种聚肌苷酸-聚胞苷酸复合物的脂质纳米佐剂及其制备方法与应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994014454A1 (fr) * 1992-12-29 1994-07-07 Entremed, Inc. Vaccins contre les sterols
US20110206758A1 (en) * 2005-12-13 2011-08-25 Pierre Vandepapeliere Vaccine Compositions Comprising a Saponin Adjuvant
WO2012088414A1 (fr) * 2010-12-23 2012-06-28 Ludwig Institute For Cancer Research Ltd. Formulation liposomale de céramides non-glycosidiques et leurs utilisations

Family Cites Families (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4186183A (en) 1978-03-29 1980-01-29 The United States Of America As Represented By The Secretary Of The Army Liposome carriers in chemotherapy of leishmaniasis
US4302459A (en) 1980-03-19 1981-11-24 The United States Of America As Represented By The Secretary Of The Army Liposome carriers in leishmaniasis chemotherapy with 8-aminoquinoline derivatives
US5916588A (en) 1984-04-12 1999-06-29 The Liposome Company, Inc. Peptide-containing liposomes, immunogenic liposomes and methods of preparation and use
US5231112A (en) 1984-04-12 1993-07-27 The Liposome Company, Inc. Compositions containing tris salt of cholesterol hemisuccinate and antifungal
US4684479A (en) 1985-08-14 1987-08-04 Arrigo Joseph S D Surfactant mixtures, stable gas-in-liquid emulsions, and methods for the production of such emulsions from said mixtures
US6759057B1 (en) 1986-06-12 2004-07-06 The Liposome Company, Inc. Methods and compositions using liposome-encapsulated non-steroidal anti-inflammatory drugs
CN1093255A (zh) * 1986-06-17 1994-10-12 恩特雷麦德有限公司 抗甾醇疫苗
IE60901B1 (en) 1986-08-21 1994-08-24 Vestar Inc Improved treatment of systemic fungal infections with phospholipid particles encapsulating polyene antifungal antibiotics
US6406713B1 (en) 1987-03-05 2002-06-18 The Liposome Company, Inc. Methods of preparing low-toxicity drug-lipid complexes
US5057540A (en) 1987-05-29 1991-10-15 Cambridge Biotech Corporation Saponin adjuvant
AU598958B2 (en) 1987-11-12 1990-07-05 Vestar, Inc. Improved amphotericin b liposome preparation
US5888519A (en) 1988-06-02 1999-03-30 The United States Of America As Represented By The Secretary Of The Army Encapsulated high-concentration lipid a compositions as immunogenic agents to produce human antibodies to prevent or treat gram-negative bacterial infections
US5215680A (en) 1990-07-10 1993-06-01 Cavitation-Control Technology, Inc. Method for the production of medical-grade lipid-coated microbubbles, paramagnetic labeling of such microbubbles and therapeutic uses of microbubbles
PL170980B1 (pl) * 1992-06-25 1997-02-28 Smithkline Beecham Biolog Szczepionka PL PL PL PL PL PL PL
GB9320668D0 (en) * 1993-10-07 1993-11-24 Secr Defence Liposomes containing particulare materials
UA56132C2 (uk) * 1995-04-25 2003-05-15 Смітклайн Бічем Байолоджікалс С.А. Композиція вакцини (варіанти), спосіб стабілізації qs21 відносно гідролізу (варіанти), спосіб приготування композиції вакцини
US6846489B1 (en) * 1995-04-25 2005-01-25 Smithkline Beecham Biologicals S.A. Vaccines containing a saponin and a sterol
JP2001501173A (ja) 1996-08-23 2001-01-30 アルザ コーポレイション シスプラチン化合物を含有するリポソーム
AU4907897A (en) 1996-10-11 1998-05-11 Sequus Pharmaceuticals, Inc. Therapeutic liposome composition and method
US6056973A (en) 1996-10-11 2000-05-02 Sequus Pharmaceuticals, Inc. Therapeutic liposome composition and method of preparation
NZ336538A (en) 1996-10-15 2001-11-30 Liposome Co Inc N-acyl phosphatidylethanolamine-mediated liposomal drug delivery
US6231859B1 (en) 1996-12-02 2001-05-15 Aquila Biopharmaceuticals, Inc. Saponin adjuvant compositions
GB9817052D0 (en) * 1998-08-05 1998-09-30 Smithkline Beecham Biolog Vaccine
WO2008109398A1 (fr) * 2007-03-01 2008-09-12 The Catholic University Of America Bactériophage t4 lié à un substrat
GB0910046D0 (en) * 2009-06-10 2009-07-22 Glaxosmithkline Biolog Sa Novel compositions

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994014454A1 (fr) * 1992-12-29 1994-07-07 Entremed, Inc. Vaccins contre les sterols
US20110206758A1 (en) * 2005-12-13 2011-08-25 Pierre Vandepapeliere Vaccine Compositions Comprising a Saponin Adjuvant
WO2012088414A1 (fr) * 2010-12-23 2012-06-28 Ludwig Institute For Cancer Research Ltd. Formulation liposomale de céramides non-glycosidiques et leurs utilisations

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
GARCON NATHALIE ET AL: "Recent clinical experience with vaccines using MPL- and QS-21-containing adjuvant systems", EXPERT REVIEW OF VACCINES, EXPERT REVIEWS LTD, GB, vol. 10, no. 4, 1 April 2011 (2011-04-01), pages 471 - 486, XP009176124, ISSN: 1744-8395, DOI: 10.1586/ERV.11.29 *
See also references of EP3122380A1 *
ZOLTAN BECK ET AL: "Detection of liposomal cholesterol and monophosphoryl lipid A by QS-21 saponin and Limulus polyphemus amebocyte lysate", BIOCHIMICA ET BIOPHYSICA ACTA (BBA) - BIOMEMBRANES, vol. 1848, no. 3, 1 March 2015 (2015-03-01), pages 775 - 780, XP055194486, ISSN: 0005-2736, DOI: 10.1016/j.bbamem.2014.12.005 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017153993A1 (fr) * 2016-03-09 2017-09-14 Technion Research And Development Foundation Ltd. Formulations liposomales et leurs procédés d'utilisation en agriculture
IL261655A (en) * 2016-03-09 2018-10-31 Technion Res & Development Found Ltd Liposomal compositions and their uses in agriculture
CN109152361A (zh) * 2016-03-09 2019-01-04 技术研究及发展基金有限公司 脂质体制剂及在农业中使用该脂质体制剂的方法
EP3615005A4 (fr) * 2017-04-28 2021-03-10 The Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc. Compositions et méthodes d'administration de vaccins
US11583578B2 (en) 2017-04-28 2023-02-21 The Henry M. Jackson Foundation For The Advancement Of Military Medicine, Inc. Plasmodium falciparum recombinanr xiexumapoeozoite protein compositions and method for vaccine delivery
WO2019051149A1 (fr) * 2017-09-08 2019-03-14 Infectious Disease Research Institute Formulations liposomales comprenant de la saponine et procédés d'utilisation
WO2023175454A1 (fr) 2022-03-14 2023-09-21 Pfizer Inc. Procédés de production d'adjuvant

Also Published As

Publication number Publication date
AU2015236106A1 (en) 2016-10-20
RU2016141622A3 (fr) 2018-10-26
SG11201607396WA (en) 2016-10-28
US10434167B2 (en) 2019-10-08
EP3122380A1 (fr) 2017-02-01
RU2016141622A (ru) 2018-04-25
SG10201808315UA (en) 2018-10-30
JP2017515889A (ja) 2017-06-15
CN107124869B (zh) 2022-04-01
KR20170007242A (ko) 2017-01-18
MX2016012168A (es) 2017-04-25
US20170182152A1 (en) 2017-06-29
CA2943190A1 (fr) 2015-10-01
JP6608422B2 (ja) 2019-11-20
CN107124869A (zh) 2017-09-01
KR102242875B1 (ko) 2021-04-20

Similar Documents

Publication Publication Date Title
US10434167B2 (en) Non-toxic adjuvant formulation comprising a monophosphoryl lipid A (MPLA)-containing liposome composition and a saponin
Beck et al. Differential immune responses to HIV-1 envelope protein induced by liposomal adjuvant formulations containing monophosphoryl lipid A with or without QS21
De Serrano et al. Liposomal vaccine formulations as prophylactic agents: design considerations for modern vaccines
Chatzikleanthous et al. Lipid-based nanoparticles for delivery of vaccine adjuvants and antigens: toward multicomponent vaccines
EP0626169B1 (fr) Forme de dosage comprenant un antigène et un sel d'un dérivé organoacide d'un stérol
Gregoriadis Immunological adjuvants: a role for liposomes
Beck et al. Immune response to antigen adsorbed to aluminum hydroxide particles: Effects of co-adsorption of ALF or ALFQ adjuvant to the aluminum-antigen complex
JP2828391B2 (ja) オリゴ糖を表面に有するリポソーム
US5026557A (en) Adjuvant composition
KR20190025822A (ko) 페길화된 리포솜 및 이의 용도
US10588963B2 (en) Methods for providing adjuvanted virosomes and adjuvanted virosomes obtainable thereby
US6117449A (en) Method for inducing a systemic immune response to a hepatitis antigen
EA024074B1 (ru) Водная адъювантная композиция, содержащая неионный агент, обеспечивающий изотоничность, иммуногенная композиция и набор на её основе и способ приготовления иммуногенной композиции
WO1993017702A1 (fr) Vaccin oral
KR20140130664A (ko) 외피 바이러스에 의해 야기되는 질병의 치료를 위한 시알로어드히신의 억제제들
US10695424B2 (en) Method of making a liposome composition
US20170165358A1 (en) Methods for enhancing the immunostimulation potency of aluminum salt-absorbed vaccines
JPH05339169A (ja) 経口ワクチン
WO1990001948A1 (fr) Vaccin contre la grippe et nouveaux adjuvants
Gregoriadis mmunoiogical adjuvants: a role for
WO1999000142A1 (fr) Methode pour provoquer une reponse immunitaire systemique a un antigene de l'hepatite ou du vih

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15716264

Country of ref document: EP

Kind code of ref document: A1

REEP Request for entry into the european phase

Ref document number: 2015716264

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2015716264

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2943190

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 15127081

Country of ref document: US

WWE Wipo information: entry into national phase

Ref document number: MX/A/2016/012168

Country of ref document: MX

ENP Entry into the national phase

Ref document number: 20167026025

Country of ref document: KR

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 2017502920

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2015236106

Country of ref document: AU

Date of ref document: 20150325

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 2016141622

Country of ref document: RU

Kind code of ref document: A

REG Reference to national code

Ref country code: BR

Ref legal event code: B01A

Ref document number: 112016021644

Country of ref document: BR

ENP Entry into the national phase

Ref document number: 112016021644

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20160921